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Results for “Local Lymph Node Assay”

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PubMed1984

Effects of tumor growth on interleukins and circulating immune complexes. Mechanisms of immune unresponsiveness.

This study delineates the temporal relationship between immune complex formation and tumor growth, and provides one possible explanation for host immunosuppression during tumor growth. The authors have studied serial circulating immune complex (CIC) levels and interleukin (IL) elaboration by peripheral blood cells (IL-1 production by adherent mononuclear cells [AMC]; and IL-2 generation by peripheral blood mononuclear cells [PBMC]) during the growth of syngeneic tumor isografts in an inbred rat model. Male Wistar/Furth (W/Fu) rats were injected, subcutaneously (SC) with 2 X 10(6) W163 ( a dimethylhydrazine [DMH]-induced colon adenocarcinoma) cells into their hind limbs. Serial CIC levels, (measured by the antigen nonspecific polyethylene glycol turbidity assay) and IL-1 and IL-2 production were measured before isografting and weekly thereafter. Progressive local tumor growth occurred for 3 weeks followed by regional lymph node metastases during the fourth week. During local tumor growth, there was a progressive rise in CIC levels (123% rise compared with baseline value; P less than 0.05) which correlated with a fall in both IL-1 and IL-2 generation (r = -0.768). At the time of regional metastasis, the mean CIC levels declined, and there was a further significant decrease in IL production (IL-1 = 0.9% and IL-2 = 10% of controls in tumor bearers). These results show that progressive tumor growth results in decreased IL production by host PBC, and suggest that CIC may be involved in regulating IL generation.

Adenocarcinoma↗

PubMed1992

Cell-mediated immune response in rotavirus-infected calves: leucocyte migration inhibition assay.

The cell-mediated immune (CMI) response was determined in rotavirus-infected calves by leucocyte migration inhibition assay with blood, spleen, mesenteric lymph node and intestinal lymphocytes. The inhibition of migration was more prominent in intestinal and mesenteric lymph node lymphocytes than in spleen and blood. In rotavirus-infected calves, the assay indicated the presence of CMI response which was more prominent at the local site of infection.

Animals↗

PubMed2006

Cytokine modulation of the innate immune response in feline immunodeficiency virus-infected cats.

BACKGROUND: In vitro data suggest that innate immune function in human immunodeficiency virus type 1-infected patients is compromised; however, in vivo studies are lacking. Feline immunodeficiency virus (FIV) infection in cats provides an excellent model to explore innate immune function in vivo. The innate response against Listeria monocytogenes is well understood, making it a useful immune probe. METHODS: Recombinant L. monocytogenes carrying eukaryotic expression plasmids for feline tumor necrosis factor (TNF)- alpha , interleukin (IL)-10, interferon (IFN)- gamma , and IL-15 were created to determine whether specific cytokines would modulate innate immune function. L. monocytogenes was delivered subcutaneously, and local lymph nodes were evaluated for size, cell subpopulations, and L. monocytogenes burden. Two months later, memory responses were evaluated by IFN- gamma enzyme-linked immunospot assay. RESULTS: FIV-positive cats had significantly less lymph-node enlargement and a greater L. monocytogenes burden than FIV-negative control cats. TNF- alpha improved listericidal activity in FIV-negative control cats but not in FIV-positive cats, whereas IL-10 modestly reduced function in FIV-negative control cats. IFN- gamma improved memory responses but not clearance of L. monocytogenes. IL-15 improved innate function in FIV-positive cats and increased the percentage of natural killer cells. CONCLUSIONS: Lentivirus infection impairs innate immune function in vivo, and IL-15 can significantly restore function. We hypothesize that altered dendritic-cell function and increased regulatory T cell activity may underlie the innate immune defect in HIV infection.

Animals↗

PubMed1980

Cellular immunity during pregnancy. I. Proliferative and cytotoxic reactivity of paraaortic lymph nodes.

The possibility of changes in immune reactivity during pregnancy was studied by measuring cellular immunity in vitro of the paraaortic (PA) lymph nodes, which drain the uterus in pregnant mice. The proliferation of PA lymph node cells from primiparous pregnant C57Bl/6J mice, in mixed lymphocyte cultures (MLC) against alloantigens, was lower in magnitude, but had the same kinetics, as the response of virgins. This was observed in syngeneic and allogeneic (to DBA/2J) pregnancies, and using the paternal as well as third party allogeneic stimulators. The response was depressed by day 8 of gestation and returned to normal two days after delivery. The decrease was not due to an active suppressor mechanism, as assayed by mixing experiments. Irradiation (1500R) of the lymphocytes from pregnant mice, prior to mixing with cells from virgins, did not reveal the presence of a radioresistant suppressor cell. In contrast to the MLC results, no differences were found between lymphocytes from pregnant and virgin mice in their ability to develop MLC-generated cell-mediated lympholysis (CML) against alloantigens, as measured in a chromium release assay. The PA lymph node cells from pregnant animals bearing allogeneic fetuses also did not show evidence of in vivo sensitization to the paternal alloantigens. Therefore, the local nodes draining the uterus from primiparous pregnant animals bearing syngeneic or allogeneic fetuses show a nonspecific decrease of MLC proliferation while retaining the capacity to generate normal CML activity.

Animals↗

PubMed1981

Distribution of ethambutol in primate tissues and cells.

Ethambutol (EMB) concentrations that kill Mycobacterium tuberculosis in vitro accumulated in squirrel monkey tissues and cells known to be sites of tubercular infections. After oral administration of a clinically relevant 25 mg/kg dose, the whole-body distribution and intracellular localization of EMB were studied by radioautography. Tissue concentrations of drug were assayed by radiochemical and microbiological methods. The EMB was distributed rapidly and widely to most body tissues including lung and localized within pulmonary alveolar and axillary lymph node macrophages. The EMB in lung at 2 and 5 h after drug administration was markedly higher than the corresponding plasma concentrations and exceeded concentrations that are bactericidal in vitro for tubercle bacilli. These observations may help explain the early bactericidal activity of EMB in humans. Similarities in plasma and tissue concentrations of the drug in both species suggest the usefulness of the squirrel monkey as a model for the use of EMB in humans.

Administration, Oral↗

PubMed2004

Clinical trials update of the European Organization for Research and Treatment of Cancer Breast Cancer Group.

The present clinical trial update consists of a review of two of eight current studies (the 10981-22023 AMAROS trial and the 10994 p53 trial) of the European Organization for Research and Treatment of Cancer Breast Cancer Group, as well as a preview of the MIND-ACT trial. The AMAROS trial is designed to prove equivalent local/regional control for patients with proven axillary lymph node metastasis by sentinel node biopsy if treated with axillary radiotherapy instead of axillary lymph node dissection, with reduced morbidity. The p53 trial started to assess the potential predictive value of p53 using a functional assay in yeast in patients with locally advanced/inflammatory or large operable breast cancer prospectively randomised to a taxane regimen versus a nontaxane regimen.

Breast Neoplasms↗

PubMed1992

Popliteal lymph node enlargement induced by procainamide.

The ability of procainamide (PA) to induce primary local popliteal lymph node (PLN) reactions has been investigated. We employed an in vitro drug-metabolizing system, based on the hypothesis that the negative result for PA in a PLN assay (PLNA) was due to insufficient metabolizing activity at the reaction site. PA was incubated previously in vitro with the S-9 mixture derived from rat liver. The reactants were ultrafiltered in order to eliminate high molecular-weight molecules, and then the low molecular-weight fraction was subcutaneously (s.c.) injected into the hind footpad of mice. PLN reactions were assessed by weighing the popliteal lymph node of the injected side. The reactants of more than 5 mg of PA and S-9 mixture induced PLN enlargement in C3H/He mice 8 days after injection. BALB/c and C57BL/6 mice were also susceptible to PLN reaction in response to the reactants of PA (10 mg) and S-9 mixture. PLN reactions to PA were induced 2 days after the injection and sustained until 18 days. Contact of PA with the S-9 mixture for 30 min at 37 degrees C was sufficient to induce PLN enlargement. However, contact for 24 h reduced the peak reaction. On the other hand, PA which had not been incubated with the S-9 mixture and acetylprocainamide (acetylPA) gave little or no reaction. Local PNA has not been considered to be suitable for the detection of drugs with the potential to induce immune disorders in cases where a metabolite contributes to the adverse reaction. However, the employment of an in vitro drug-metabolizing system may overcome that defect.

Animals↗

PubMed1989

Progesterone receptors in carcinomas of the upper aerodigestive tract.

This study had three major goals: (1) to vigorously verify the presence of progesterone receptors in squamous cell carcinoma of the upper aerodigestive tract (HN-SCC). Antiprogesterone receptor monoclonal antibodies revealed a distinct band at approximately 120 kilodaltons in samples taken from two of four patients with HN-SCC. These results illustrate that progesterone receptor in HN-SCC has the same molecular weight as progesterone receptor in normal human uterus and human breast cancer. Steroid specificity and saturability results support the evidence that it is true progesterone receptors that are measured and not other receptors or sex steroid-binding globulins; (2) to confirm the biochemical function of progesterone receptors in HN-SCC by assessing the binding of progesterone receptor to acceptor sites on chromosomes in the nucleus; and (3) to establish the clinical significance of progesterone receptor measurement. Patients with positive assays were more likely to be free of disease a mean of 6 months after resection. We used logistic regression to account for site of primary disease, grade of tumor, and stage of disease. This logistic regression was significant with a p = 0.014. Patients with a binding index greater than 2 (19 of 73 patients) were 4.34 times more likely to be free of disease than patients with negative assays.

Adult↗

PubMed2002

Transplantation of neural stem cells in a rat model of stroke: assessment of short-term graft survival and acute host immunological response.

The use of progenitors and stem cells for neural grafting is promising, as these not only have the potential to be maintained in vitro until use, but may also prove less likely to evoke an immunogenic response in the host, when compared to primary (fetal) grafts. We investigated whether the short-term survival of a grafted conditionally immortalised murine neuroepithelial stem cell line (MHP36) (2 weeks post-implantation, 4 weeks post-ischaemia) is influenced by: (i) immunosuppression (cyclosporin A (CSA) vs. no CSA), (ii) the local (intact vs. lesioned hemisphere), or (iii) global (lesioned vs. sham) brain environment. MHP36 cells were transplanted ipsi- and contralateral to the lesion in rats with middle cerebral artery occlusion (MCAo) or sham controls. Animals were either administered CSA or received no immunosuppressive treatment. A proliferation assay of lymphocytes dissociated from cervical lymph nodes, grading of the survival of the grafted cells, and histological evaluation of the immune response revealed no significant difference between animals treated with or without CSA. There was no difference in survival or immunological response to cells grafted ipsi- or contralateral to the lesion. Although a local upregulation of immunological markers (MHC class I, MHC class II, CD45, CD11b) was detected around the injection site and the ischaemic lesion, these were not specifically upregulated in response to transplanted cells. These results provide evidence for the low immunogenic properties of MHP36 cells during the initial period following implantation, known to be associated with an acute host immune response and ensuing graft rejection.

Animals↗

PubMed1986

A quantitative lymphocyte localization assay.

Multiple lesions (up to 100 sites) were induced in the skin of sheep using either allogeneic lymphocytes or, in BCG-sensitized animals, tuberculin. Cells recovered from an indwelling lymph catheter draining a prescapular lymph node were labeled with 111-indium, returned to venous blood, and allowed to circulate for 3 hours. Sheep were killed, and the skin lesions and lymph nodes were removed and counted in a gamma spectrometer. High levels of radioactivity (up to 38,000 cpm/lesion) were recovered from lesions, and only a few hundred cpm were recovered from comparable normal skin sites. Dose-response relationships and time kinetics were demonstrated for these lesions, and the radioactivity on blood and lymph cells was measured. The contribution of cell-free radioactivity was negligible. Using replicate injection sites, analytical, internally controlled studies can now be initiated to study the induction, promotion, and suppression of lymphocyte traffic.

Animals↗

PubMed1984

Allogeneic reactivity of maternal lymphoid cells during the course of gestation. Modifications and sex differences in a local GVH assay.

The immune reactivity of lymphoid cells from pregnant mice was studied during the course of pregnancy in primiparous and multiparous animals either " isopregnant " (male and female of same strain) or " allopregnant " (male and female differing at H-2), using a local GVH assay (CBA lymphoid cells injected into (CBA X A/J)F1 recipients). The findings were as follows: The lymphoid cell number in the para-aortic lymph nodes ( PALN ) was increased at all stages of gestation. The peak occurred in the 2nd week in primiparity and as early as 60 h after fertilization in multiparity. PALN cell alloreactivity was weak at the beginning and higher than normal in the third week of pregnancy. Spleen cell alloreactivity was increased in the second week and decreased in the third week in primiparous compared with multiparous animals. Anti-paternal alloreactivity exhibited by spleen cells of allogestation was decreased (as compared to cells of isogestation ) especially in primiparous mice, particularly in the third week. At this time, the anti-paternal alloreactivity of PALN cells was increased. The influence of the recipient's sex on GVHR intensity was reversed when the cells were obtained from a pregnant donor, becoming stronger in male compared with female hybrids.

Animals↗

PubMed1999

Clinical significance of the detection of thymidine phosphorylase activity in esophageal squamous cell carcinomas.

In this study, we obtained fresh samples of tissue (tumors, adjacent normal mucosas, and metastatic lymph nodes) from 64 patients (metastatic lymph nodes were obtained from 16 patients) with esophageal cancer who underwent esophagectomy between 1993 and 1998. The thymidine phosphorylase (dThdPase) levels of tumors as determined by the ELISA method were compared with clinicopathological findings of tumors and were evaluated for their usefulness as a prognostic parameter for these patients. The dThdPase levels of tumors (221 +/- 21 U/mg protein) and metastatic lymph nodes (253 +/- 51 U/mg protein) were significantly higher than the dThdPase level of normal mucosas (53 +/- 7 U/mg protein, p < 0. 001). The dThdPase levels of tumors did not correlate with the histopathological grading of tumors, the depth of tumor invasion, and lymph node metastasis. The 3-year survival rate of 32 patients with a high level of dThdPase (42%) was not different from that of 32 patients with a low level of dThdPase (52%, p = 0.267). Moreover, the dThdPase level of tumors was not an independent prognostic factor for patients with esophageal cancer. These findings suggest that the level of dThdPase activity in esophageal cancers may not correlate with tumor progression and may not be a useful prognostic marker for patients with esophageal squamous cell carcinoma.

Aged↗

PubMed1997

The alloreactivity in the popliteal lymph node (PLN) assay is regulated by malononitrilamides (MNAs).

Malononitrilamides (MNA 279 and MNA 715) represent a new class of low molecular weight immunosuppressants and belong to the derivatives of the primary metabolite of leflunomide A7771726. They have been shown to prevent and reverse established acute allograft rejection and effectively prolong xenograft survival, and have also been found to be potent inhibitors of B- and T-cell mediated autoimmune processes. The MNAs mediate their effects by binding specifically to dehydro orotate-dehydrogenase (DHODH) and inhibiting de novo pyrimidine biosynthesis, thereby blocking T- and B-cell proliferation and strongly suppressing the IgM and IgG antibody production. In this study we evaluated the effects of MNA 279 and MNA 715 on the in vivo lymphoproliferation that occurs after challenge with allogeneic cells in a local graft-versus-host (GvH) reaction in Lewis x Brown-Norway (LBN) F1-hybrid rats by measuring the enlargement of the PLN draining the site of allogeneic cell injection. Oral administration of one of the two MNAs (7.5 to 50 mg/kg) on day 0, dose-dependently prevented the localized lymphoproliferative response and suppressed the lymph node hyperplasia. The MNAs even acted therapeutically when they were given during an ongoing alloreactivity as late as day 4 or 5 after challenge. Consistent with the mode of action that MNAs inhibit de novo pyrimidine biosynthesis, a complete reversal of the immunosuppression on the lymphoproliferation in vivo was attempted in this protocol by addition of exogenous uridine during days 0 to 5. These data suggest that MNA 279 and MNA 715 mediate their antiproliferative and immunosuppressive effects in the PLN-assay in vivo by decreasing the activity of DHODH in the lymph node cells and thereby inhibiting pyrimidine biosynthesis.

Alkynes↗

PubMed1986

Selective chemotaxis of subsets of B lymphocytes from gut-associated lymphoid tissue and its implications for the recruitment of mucosal plasma cells.

As they differentiate, precursor cells from the gut-associated lymphoid tissue are known to travel via the lymphatic system to the blood and then preferentially to home to various mucosal and exocrine sites such as the lamina propria of the gut and the lactating mammary gland, where they give rise to IgA-secreting plasma cells. The present study, directed at the mechanism by which the circulating precursors of mucosal IgA plasma cells selectively lodge in characteristic locations, explored the hypothesis that such homing is due to a locally produced chemotactic factor and that milk might be a source of such a factor. Subsets of lymphocytes bearing particular surface markers and purified by panning from lymph nodes of mice were examined in a micropore chemotaxis assay to search for the presence of chemotactic activity in mouse milk. The globulin fraction of whey was shown to contain a nondialyzable factor that is chemotactic for IgA (and also IgG)-positive lymphocytes when these are obtained from mesenteric lymph nodes as a source of mucosal-associated lymphoid tissue. Lymphocytes from peripheral lymph nodes, nonmucosal associated, were unaffected as were surface IgM-positive lymphocytes and T lymphocytes obtained from mesenteric nodes. Chemotactic activity for IgA lymphocytes was undetectable in mouse serum. The data are consistent with the idea that precursors of mucosal IgA plasma cells home to mucosal and exocrine sites in response to a specific chemotactic factor elaborated by local differentiated epithelial cells.

Animals↗

PubMed1982

[Carcino-embryonic antigen assay in cancer of the upper respiratory and digestive tracts].

Radioimmunological assay of carcino-embryonic antigen (CEA) was performed in 238 patients with cancer of the upper respiratory and digestive tracts after a minimum follow-up period of 3 years. Secretion of CEA was observed in only 1 out of 3 cases, levels, always being low and only slightly influenced by the localization or size of the tumor, and the condition of the lymph nodes. Initial levels are of no prognostic significance. The appearance of high levels during the course of the disease corresponds to shorter survival, but as it does not precede clinical evidence of recurrence or metastases this assay cannot play a role as a tumor marker.

Carcinoembryonic Antigen↗

PubMed2001

Germinal center reaction in the joints of mice with collagen-induced arthritis: an animal model of lymphocyte activation and differentiation in arthritis joints.

OBJECTIVE: To establish an animal model and provide a basis for investigating the role of germinal center (GC) reaction in autoimmune arthritis. METHODS: DBA/1 mice were immunized with bovine type II collagen (CII) to elicit collagen-induced arthritis (CIA). Sections of arthritic joints were examined by in situ immunohistochemical studies, and purified cells from affected joints were subjected to flow cytometry analysis. RESULTS: De novo GC reaction was induced in the arthritic joints of male DBA/1 mice by immunization with bovine CII. In comparison with GCs formed in lymphoid tissues, such as spleen and lymph nodes, we found that these GCs formed in the joint tissues of mice with CIA were morphologically typical, as determined by immunohistologic and flow cytometric assays. CONCLUSION: The local immune responses in murine CIA induced ectopic GC formation, as observed in the synovial tissues of patients with rheumatoid arthritis. This system will allow for the first time the direct study of the role of the GC reaction in autoimmune arthritis in an animal model.

Animals↗

PubMed2003

Bacterial translocation up-regulates GTP-cyclohydrolase I in mesenteric vasculature of cirrhotic rats.

In cirrhosis, arterial vasodilation and the associated hemodynamic disturbances are most prominent in the mesenteric circulation, and its severity has been linked to bacterial translocation (BT) and endotoxemia. Synthesis of nitric oxide (NO), the main vasodilator implicated, is dependent on the essential cofactor tetrahydrobiopterin (BH(4)). The key enzyme involved in BH(4) synthesis is GTP-cyclohydrolase I (GTPCH-I), which is stimulated by endotoxin. Therefore, we investigated GTPCH-I activity and BH(4) biosynthesis in the mesenteric vasculature of cirrhotic rats with ascites, as well as their relationship with BT and endotoxemia, serum NO, and mean arterial pressure (MAP). GTPCH-I activity and BH(4) content in mesenteric vasculature was determined by high-performance liquid chromatography. BT was assessed by standard bacteriologic culture of mesenteric lymph nodes (MLNs). Serum endotoxin was measured by a kinetic turbidimetric limulus amebocyte lysate assay, and serum NO metabolite (NOx) concentrations were assessed by chemiluminescence. BT was associated with local lymphatic and systemic appearance of endotoxin and was accompanied by increases in serum NOx levels. GTPCH-I activity and BH(4) content in mesenteric vasculature were both increased in animals with BT and correlated significantly (r = 0.69, P <.01). Both GTPCH-I activity and BH(4) levels significantly correlated with serum endotoxin and NOx levels (r = 0.69 and 0.54, 0.81 and 0.53, P <.05). MAP (a marker of systemic vasodilatation) correlated with endotoxemia (r = 0.58, P <.03) and with GTPCH-I activity (r = 0.69, P <.01). In conclusion, in cirrhotic animals BT appears to lead to endotoxemia, stimulation of GTPCH-I, increased BH(4) synthesis, and further enhancement of vascular NO production that leads to aggravation of vasodilatation.

Animals↗

PubMed1981

Quantitative oestradiol receptor values in primary breast cancer and response of metastases to endocrine therapy.

Quantitative values if oestradiol receptor were determined in the primary breast cancer of 526 patients, 106 of whom have so far required hormonal therapy for metastatic disease (premenopausal patients-oophorectomy; postmenopausal patients-tamoxifen 20 mg twice daily). The rate of response to treatment was significantly higher in patients with receptor-positive primary cancers, and the likelihood of response further increased in proportion to the measured receptor concentration. Preliminary data also suggest that patients with high receptor values in the primary tumour will enjoy a longer remission than those with low measured values. Assays of oestradiol receptor were also carried out on biopsy specimens of accessible metastases, largely skin and lymph-node recurrences, in 24 patients who were treated mainly with local measures. Receptor status was the same in both the primary and the secondary tumour in only 18 patients.

Adult↗